N,N'-diacetylhydrazine

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: - meets generally accepted scientific principles - Documentation insufficient for thorough assessment

Data source

Reference

Reference 1

Reference Type:

publication

Title:

Mutagenicity and carcinogenicity of mono- and diacetyl hydrazine.

Author:

Bhide SV, Bhalerao EB, Sarode AV, Maru GB.

Year:

1984

Bibliographic source:

Cancer Lett. 1984 Jun;23(2):235-40.

Reference 2

Reference Type:

publication

Title:

The in vivo micronucleus assay in mammalian bone marrow and peripheral blood. A report of the U.S. Environmental Protection Agency Gene-Tox Program.

Author:

Mavournin KH, Blakey DH, Cimino MC, Salamone MF, Heddle JA.

Year:

1990

Bibliographic source:

Mutat Res. 1990 Jul;239(1):29-80.

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

: 4 animals per dose, 2 treatments at an interval of 24 h, single sampling 6 h after last treatment.

GLP compliance:

no

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-diacetylhydrazine

EC Number:

221-576-1

EC Name:

N,N'-diacetylhydrazine

Cas Number:

3148-73-0

Molecular formula:

C4H8N2O2

IUPAC Name:

N'-acetylacetohydrazide

Details on test material:

- Name of test material (as cited in study report): diacetyl hydrazine (DAH)
- Physical state:
• diacetylhydrazine: not reported
- Analytical purity:
• diacetylhydrazine: not reported
- Impurities (identity and concentrations):
• diacetylhydrazine: not reported
- Composition of test material, percentage of components:
• diacetylhydrazine: not reported
- Provider: Ega Chemie, Germany (nowadays Sigma-Aldrich)

Test animals

Species:

mouse

Strain:

Swiss

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS

- Age at study initiation: 6 - 8 weeks
- Weight at study initiation: 20 - 30 g

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

- Vehicle(s)/solvent(s) used: none
- Justification for choice of solvent/vehicle: N/A

Details on exposure:

APPLICATION
- ip injection of pure diacetylhydrazine: at 2400 mg/kg bw/d

Duration of treatment / exposure:

- twice, at an interval of 24 h

Frequency of treatment:

- twice, at an interval of 24 h

Post exposure period:

not applicable animals sacrificed 6 h after last dosing

Doses / concentrations

Remarks:

Doses / Concentrations:
2400 mg/kg bw/d of diacetylhydrazine
Basis:
other: actual injected

No. of animals per sex per dose:

4 males per dose

Control animals:

other: yes, concurrent treatment with 0.2 mg water/animal

Positive control(s):

Monoacetyl hydrazine was tested in parallel and showed positive results. It can be regarded as a positive control.

Examinations

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION:
- Based on LD50 values of 3000 mg diacetyl hydrazine/kg bw in mice

DETAILS OF SLIDE PREPARATION:
- bone marrow preparations
- stained in May-Grunwald Giemsa
- mounted in Euparol

METHOD OF ANALYSIS:
- 4000 polychromatic and normochromatic erythrocytes scored per animal

OTHER:

Evaluation criteria:

not reported

Statistics:

Results are mean ± S.E. of 4 animals in each group.

Results and discussion

Test results

Sex:

male

Genotoxicity:

negative

Remarks:

for diacetylhydrazine

Toxicity:

no effects

Remarks:

, based on an unaffected ratio of polychromatic cells/normochromatic cells as compared to negative control

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- no range finding study reported

RESULTS OF DEFINITIVE STUDY
- acetylhydrazine: significant increase in the frequency of micronuclei in the polychromatic erythrocyte.
- diacetylhydrazine: no significant increase in the frequency of micronuclei in the polychromatic erythrocyte.

Any other information on results incl. tables

- Table 1: mutagenicity of DAH using the micronucleus test

No.	Compound	Dose/  animal (mg)	% Polychromatic   erythrocytes with micronucleus	P/N   ratio (a)
1	Distilled water	0.2	0.25 ± 0.05	1.0
2	DAH	48	0.36 ± 0.03	1.1

Results are expressed as mean ± S.E. of 4 animals in each group.

(a) P/N ratio, ratio of polychromatic to normochromatic cells.

*Statistically significant when compared with controls, P < 0.05.

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative
Diacetylhydrazine was tested for it's potential to cause chromosome abberations in an in vivo micronucleus test in Swiss mice 2400 mg/kg bw ip injected twice, conducted comparable to OECD 474. No adverse effects were seen for diacetylhydrazine at the tested dose. Accordingly diacetylhydrazine (DAH) is deemed negative for in vivo chromosome abberation in this test system.
Though the details reported are too limited for a solid assessment, the results are deemed relevant and sufficient as supporting information.

Executive summary:

In the study Bhide (1984) diacetylhydrazine were tested for their potential to cause chromosome abberations in an in vivo micronucleus test in Swiss mice at 2400 mg/kg bw ip injected twice, conducted comparable to OECD 474.

4 male Swiss mice per dose were used and the animal were treated twice at an interval of 24 h. Sampling was conducted once, 6 h after the last treatment. Bone marrow preparations were made, stained in May-Grunwald Giemsa and mounted in Euparol. 4000 polychromatic and normochromatic erythrocytes were scored per animal.

No adverse effects were seen for diacetylhydrazine at the tested dose. Accordingly diacetylhydrazine (DAH) is deemed negative for in vivo chromosome abberation in this test system.

Though the details reported are too limited for a solid assessment, the results are deemed relevant and sufficient as supporting information.